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According to our previous investigation the total methanol extract from Graptopetalum paraguayense E. Walther demonstrates a significant inhibitory effect on herpes simplex virus type 1 (HSV-1). To clarify what causes this inhibitory activity on HSV-1, a metabolic profile of the plant was performed. Three main fractions: non-polar substances, polar metabolites and phenolic compounds were obtained and gas chromatography–mass spectrometry (GC-MS) analysis was carried out. Since it is well known that phenolic compounds show a significant anti-herpes effect and that viral DNA polymerase (DNApol) appears to play a key role in HSV virus replication, we present a docking and quantum-chemical analysis of the binding of these compounds to viral DNApol amino acids. Fourteen different phenolic acids found by GC-MS analyses, were used in molecular docking simulations. According to the interaction energies of all fourteen ligands in the DNApol pockets based on docking results, density functional theory (DFT) calculations were performed on the five optimally interacting with the receptor acids. It was found that hydroxybenzoic acids from phenolic fraction of Graptopetalum paraguayense E. Walther show a good binding affinity to the amino acids from the active site of the HSV DNApol, but significantly lower than that of acyclovir. The mode of action on virus replication of acyclovir (by DNApol) is different from that of the plant phenolic acids one, probably.

The current study presents results from three years experimental work focused on evaluation of climate plasticity and content of free amino acids, mono- and disaccharides and fatty acids of three soybean cultivars by using an innovative approach of long-lasting low temperature pretreatment of soybean grains before sowing. The results from field performance of plants grown from pretreated and non-treated seeds were taken based on five morphological indicators. Expression and metabolomic profiles of leaves, green seeds and mature seeds of plants grown from low temperature pretreated and not-treated seeds were assessed by real time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and gas chromatography/mass spectrometry (GC/MS). The all three years collected data from the morphological analyses, expression profiles and content of metabolites confirmed that the plants grown from low temperature pretreated seeds possess better field performance, enhance content of free amino acids, sugars and fatty acids in green and mature seeds. The obtained expression profiles of selected genes correlated with the content of amino acids and sugars. The obtained results in the present study could benefit legume crops researchers, plant breeders, farmers and processors of soybean.

Rapeseed meal is a by-product of oil producing industry which contains a high amount of protein (37–42%). To prepare protein isolates, suitable for human nutrition, the industrial rapeseed meal is often subjected to a pre-treatment with aqueous ethanol solution. The aim of the study was to investigate the phytochemical profile and bioactivity of the ethanol-wash solutes (EWS) obtained as a waste product of the main process of the protein isolate preparation from industrial rapeseed meal. Proximate composition analysis revealed that total carbohydrates were more than half of the product (60.63%). The content of hydrolysable reducing sugars was 44.13%, while glucose was 7.22%. The total phenols amounted 13.38%. EWS contained 1.63% total flavonoids and 242.05 μmol/g dry weight glucosinolates. The major components, as determined by GC–MS analysis, were sucrose, turanose and melibiose with 22.68%, 4.08% and 3.97% of total ion current (TIC) of polar compounds, respectively; cetyl alcohol (2.45% of TIC) and methyl oleate (2.08% of TIC) representing nonpolar compounds, and sinapic acid (73.71% of TIC) which was the prevailing substance of the phenolic acids identified in the EWS. The 0.2% EWS, prepared in aqueous 70% ethanol solution, exhibited 70.68% scavenging capability of DPPH radicals and 585.11 μmol Fe2+/ g EWS (dry weight) reducing capacity. The product demonstrated limited antibacterial but broad antifungal activity which was expressed against Aspergillus niger (ATCC 1015), Aspergillus flavus, Penicillium sp., Rhizopus sp. and Fusarium moniliforme (ATCC 38932) as determined by the agar-well diffusion assay. Overall, EWS exhibited bioactive capacity, which revealed its potential application as a value-added product in the food and nutraceutical industries or agriculture. The broad-spectrum antifungal activity makes the product a prospective agent for food biopreservation or plant bioprotection.

Sambucus ebulus L. (SE) fruits are used for their immunostimulation, hematopoietic and antiviral potential. Recently, we focused on analyzing the mechanism underlying SE fruit aqueous extract’s (FAE) immunomodulation and anti-inflammatory activities, with attention to its endoplasmic reticulum (ER) stress-reducing potential. J774A.1 macrophages were treated with SE FAE alone or in conditions of lipopolysaccharides (LPS) stimulation. Using GC–MS and LC–MS/MS, its phytochemical composition was analyzed. To measure transcription and protein levels, we used qPCR and Western blot, respectively. The prevailing phytochemicals in SE FAE were hydroxycinnamic acids, proanthocyanidins and anthocyanins. The content of some amino acids, organic acids, alcohols, fatty acids and esters were newly reported. Extracts exerted an immunostimulation potential by stimulating IL-6, TNFα, Ccl2, COX2 and iNOS transcription, without inducing ER stress. SE FAE suppressed the LPS-induced transcription of inflammation related genes (IL-1β, IL-6, TNFα, Ccl2, Icam-1, Fabp4, COX2, iNOS, Noxo1, IL-1ra, Sirt-1) and reduced the protein levels of iNOS, peIF2α, ATF6α and CHOP. The effects were comparable to that of salicylic acid. SE suppresses LPS-stimulated inflammatory markers on the transcription and translation levels. Targeting ER stress is possibly another mechanism underlying its anti-inflammatory potential. These findings reveal the potential of SE fruits as a beneficial therapeutic of inflammation and ER stress-related pathological conditions.

The prospect of replacing traditional chemical fertilization with organic and microorganism-based fertilization meets the current demand for more sustainable cropping systems and healthy food. In this respect, research was carried out to evaluate the effects of the factorial combination between four basil cultivars (‘Aromat de Buzau’, ‘Macedon’, ‘Cuisoare’ and ‘Serafim’) and three types of fertilization, namely chemical fertilization (with a solid chemical fertilizer), organic fertilization (with chicken manure formulate) and microorganisms’ fertilization (with microorganisms formulate), on basil yield, biochemical and physiological parameters and essential oil composition. The results showed that the biometric parameters (plant height, number of stems and leaves and leaf area) were significantly influenced by the cultivar; ‘Macedon’ obtained the highest values of plant height (64.7 cm) and number of stems (20.33) and leaves (618.3) and ‘Serafim’ the largest leaf area (4901.7 cm2 per plant), while the type of fertilization did not affect these parameters. Regarding the biomass, the influence of the cultivar was not significant on fresh biomass but was significant on dry biomass, with ‘Macedon’ showing the highest value (56.4 g·plant−1 dry biomass). The mentioned parameters were significantly influenced by the type of fertilization, with the highest values recorded with chemical fertilization. Both the cultivar and the fertilization type significantly influenced the physiological parameters (the total content of assimilatory pigments and photosynthesis). Five phenolic compounds were quantified from leaf extracts by HPLC-MS (caffeic acid, hyperoside, isoquercitrin, rutin and quercitrin). Hyperoside was identified only in ‘Macedon’, while the rest of the compounds were found in all the cultivars and varied depending on the cultivar and fertilization type. Regarding the composition of the essential oil, variation was found depending on the cultivar and fertilization type. In ‘Aromat de Buzau’, the main compounds were methyl chavicol and β-linalool; in ‘Macedon’, geranial and neral; and in ‘Cuisoare’ and ‘Serafim’, β-linalool. Moreover, the PCA showed that the ‘Serafim’ cultivar has exclusive properties compared to the other cultivars. Our results highlight that identifying the most effective interaction between genotype and fertilization type allows to optimize yield and quality targets for sweet basil.

Graptopetalum paraguayense E. Walther (GP) belongs to the Crassulaceae family and has several health benefits. In the present study we evaluated the cytotoxic, anti-Herpes Simplex virus (HSV) and antibacterial activities of a total methanol/water GP extract. The results from the cytotoxicity investigation of the tested extract showed a high cell tolerable concentration range. The GP extract demonstrated a significant inhibitory effect on the wild-type HSV-1 strain Victoria in a concentration of 0.0001 mg/mL equal to the maximal nontoxic concentration (with 97.5% protection of the cells). Regarding the acyclovir-resistant mutant HSV-2 strain PU and wild-type HSV-2 strain Bja, the antiviral activity and selectivity of the extract were very low, with EC50 values from 0.1 to 0.01 mg/mL, and the selectivity index ranging from 25 to 250, respectively. The GP extract exhibited selective inhibitory activity also on the growth of Gram-positive bacterial pathogens, with Staphylococcus aureus being most sensitive. It also prevented the formation of MRSA biofilm up to 50% at a concentration of 1.6 mg/mL and up to 90% at a concentration of 2.5 mg/mL. The anti-herpetic and antibacterial activities demonstrated in our investigation are probably related to the presence of phenols, and in particular gallic acid. The antimicrobial activity of the GP extract holds a promise for the future development of nutrition supplements based on G. paraguayense and therefore warrants further studies aimed at isolating biologically active compounds or mixtures thereof with subsequent in-depth investigation of their anti-HSV and antibacterial mechanisms of action.

Bilberries (Vaccinium myrtillus L.) belong to the Ericaceae family and have been traditionally used in folk medicine for centuries because of their high content of polyphenolic compounds. Berry polyphenols exhibit strong antioxidant properties that could have preventive and therapeutic effects for cardiovascular disease, neurodegenerative disorders, cancer, diabetes, osteoporosis, obesity. More recently, scientific interest in the leaf composition and beneficial properties has been growing, documenting that berry leaves can be considered an alternative source of bioactive substances. The main compounds in berry leaves are similar to those in berry fruits, i.e., phenolic acids, flavonols, proanthocyanidins. The leaves are one of the richest sources of chlorogenic acid. The aim of the current research was to investigate the intrapolulation variations of polyphenolic compounds in a bilberry (Vaccinium myrtillus L.) habitat in order to select the most productive plant individuals for introduction in vitro. The total polyphenols, total flavonoid content, total proanthocyanidines and antioxidant capacity were estimated by Folin–Ciocalteu’s method, aluminium nitrate method, n-butanol-HCl assay and DPPH, respectively. The total polyphenol content of bilberry was 52.37 ± 8.62 mg GAE g−1 DW with coefficient of variation (CV%) 16.45%. The radical scavenging activity of the bilberry population defined by DPPH assay was 477.18±111.32 mmol TE g−1 DW with CV – 23.33%. HPLC analyses revealed that the dominan phenolic acid of the bilberry population was chlorogenic acid (17.10 ± 3.21 mg g−1 DW). The results obtained revealed that the analyzed population of bilberry possessed significant intrapopulation variations in respect of the quantity of investigated secondary metabolites

The F-box domain is a conserved structural protein motif that most frequently interacts with the SKP1 protein, the core of the SCFs (SKP1-CULLIN-F-box protein ligase) E3 ubiquitin protein ligases. As part of the SCF complexes, the various F-box proteins recruit substrates for degradation through ubiquitination. In this study, we functionally characterized an F-box gene (MtF-box) identified earlier in a population of Tnt1 retrotransposon-tagged mutants of Medicago truncatula and its Arabidopsis thaliana homolog (AtF-box) using gain- and loss-of-function plants. We highlighted the importance of MtF-box in leaf development of M. truncatula. Protein–protein interaction analyses revealed the 2-isopropylmalate synthase (IPMS) protein as a common interactor partner of MtF-box and AtF-box, being a key enzyme in the biosynthesis pathway of the branched-chain amino acid leucine. For further detailed analysis, we focused on AtF-box and its role during the cell division cycle. Based on this work, we suggest a mechanism for the role of the studied F-box gene in regulation of leucine homeostasis, which is important for growth.

The purpose of this study was to perform phytochemical analysis of tea from Sambucus ebulus fruits concerning hydroxycinnamic acids, flavonol glucosides, stilbenes and proanthocyanidin mono-, di- and trimers content. In total, 33 compounds were identified and quantified using UPLC-DAD-ESI/MS/MS system and the results are presented in mg/g dry weight (DW). Among analyzed hydroxycinnamic acids, 5-Caffeoylquinic acid (114.17 mg/g) was most abundant, followed by 3-p-Coumaroylquinic acid (50.33 mg/g) and 3-p-Feruloylquinic acid, p-Coumaric acid glucoside and 4-p-Coumaroylquinic acid (31.36 mg/g, 29.78 mg/g and 27.70 mg/g, respectively). Flavonol glucosides were represented predominantly by Quercetin-3-O-galactoside, Quercetin-3-O-rhamnosyl-galactoside Quercetin-3-O-glucoside and Quercetin-3-O-rhamnosyl-glucoside (3.68 mg/g, 3.22 mg/g, 2.87 mg/g and 2.56 mg/g, respectively). trans-Resveratrol-3-O-glucoside, epicatechin (40.62 mg/g) and proanthocyanidin di- and –trimers (19.90 mg/g – 31.42 mg/g) also were present in the tea. ABTS cation decolorization assay revealed 1.248 mM UAE activity and the percent of DPPH radical scavenging was 14.25%, corresponding to 39.07 μM Trolox equivalents.

The purpose of the current study was to identify and quantify triterpenoids and other non-polar compounds in the leaves of three high bush blueberry cultivars (Vaccinium corymbosum L. var. Bluegold, var. Bluecrop and var. Elliott) and three natural populations of Vaccinium species (Vaccinium uliginosum L., Vaccinium myrtillus L. and Vaccinium vitis-idaea L.) by means of gas chromatography mass spectrometry (GC-MS) and high-performance liquid chromatography with diode array detector (HPLC-DAD). Metabolite profiles differed significantly among the Vaccinium species analyzed, as well as among the populations of the same species. The populations of V. vitis-idaea predominantly contained relative concentrations of phytosterols (varying between 10.48% of total ion current (TIC) and 22.29% of TIC) and almost twice the content of triterpenes (from 29.84% of TIC to 49.62% of TIC) of the other berry species investigated. The leaves of V. corymbosum varieties biosynthesized the highest relative amount of fatty acids, while the leaves of the populations of V. uliginosum had the highest relative concentrations of fatty alcohols. The results of principal component analysis (PCA) and hierarchical cluster analysis (HCA) showed that the diverse populations of each berry species analyzed differed from each other, most likely due to variations in the climatic and geographical conditions of their localities.

Sambucus ebulus L. (SE) fruits are used for their immunostimulation, hematopoietic and antiviral potential. Recently, we focused on analyzing the mechanism underlying SE fruit aqueous extract’s (FAE) immunomodulation and anti-inflammatory activities, with attention to its endoplasmic reticulum (ER) stress-reducing potential. J774A.1 macrophages were treated with SE FAE alone or in conditions of lipopolysaccharides (LPS) stimulation. Using GC–MS and LC–MS/MS, its phytochemical composition was analyzed. To measure transcription and protein levels, we used qPCR and Western blot, respectively. The prevailing phytochemicals in SE FAE were hydroxycinnamic acids, proanthocyanidins and anthocyanins. The content of some amino acids, organic acids, alcohols, fatty acids and esters were newly reported. Extracts exerted an immunostimulation potential by stimulating IL-6, TNFα, Ccl2, COX2 and iNOS transcription, without inducing ER stress. SE FAE suppressed the LPS-induced transcription of inflammation related genes (IL-1β, IL-6, TNFα, Ccl2, Icam-1, Fabp4, COX2, iNOS, Noxo1, IL-1ra, Sirt-1) and reduced the protein levels of iNOS, peIF2α, ATF6α and CHOP. The effects were comparable to that of salicylic acid. SE suppresses LPS-stimulated inflammatory markers on the transcription and translation levels. Targeting ER stress is possibly another mechanism underlying its anti-inflammatory potential. These findings reveal the potential of SE fruits as a beneficial therapeutic of inflammation and ER stress-related pathological conditions.

A comparative investigation of n-hexane soluble compounds from aerial parts of dandelion (Taraxacum officinale Weber ex F.H. Wigg.) collected during different vegetative stages was carried out. The GC-MS analysis of the n-hexane (unpolar) fraction showed the presence of 30 biologically active compounds. Phytol [14.7% of total ion current (TIC)], lupeol (14.5% of TIC), taraxasteryl acetate (11.4% of TIC), β-sitosterol (10.3% of TIC), α-amyrin (9.0% of TIC), β-amyrin (8.3% of TIC), and cycloartenol acetate (5.8% of TIC) were identified as the major components in n-hexane fraction. The unpolar fraction exhibited promising antioxidant activity – 46.7 mmol Trolox equivalents/g extract (determined by 1,1-diphenyl-2-picrylhydrazyl method). This fraction demonstrated insignificant antimicrobial activity and can be used in cosmetic and pharmaceutical industries.

The development of organic farming as a result of increasing consumer preference for organic food has led to the development and registration of new pest-control products for certified organic production. In this study, the effects of three biocontrol products containing spores and mycelium of Arthrobotrys oligospora—Artis®, Beauveria bassiana—Bora®, and Coniothyrium minitans—Öko-ni® were tested on four basil (Ocimum basilicum L.) cultivars: ‘Aromat de Buzau’, ‘Serafim’, ‘Macedon’ and ‘Cuisoare’. The application of Öko-ni® increased basil yields by 8% relative to Control. The application of Bora® increased chlorophyll content of basil leaves by 2% and the activity of photosynthesis by 66% relative to the Control. Basil essential oil (EO) content was increased by 18% with the application of Artis® and by 34% with the application of Bora® and Öko-ni®, respectively. The content of phenolic compounds analyzed by HPLC varied, caffeic acid concentration was higher in the plants treated with Öko-ni®, hyperoside, isoquercitrin and rutin concentrations were higher in those treated with Artis®, while the quercitrin content was higher in Bora®-treated plants. The two main EO constituents that were identified were linalool and methyl chavicol in ‘Aromat de Buzau’, linalool and eugenol in ‘Serafim’, neral and geranial in ‘Macedon’, also linalool and eugenol in ‘Cuisoare’. The investigated myco-biocontrol products had positive effects on basil fresh biomass and EO content and also influenced the content of phenolic compounds.

The Climate Change is one of the major problems in the entire world. The limiting influence of harmful factors as climate and other environmental changes, it will be crucial to breed new varieties for improved resistance to abiotic and biotic stresses. Climate Change can be tackled through plant breeding and better use of Plant Genetic Resources. Generally, plant breeding is the art and science of genetically improving plants for the benefit of humankind. It is important to mention potential contribution of organically managed systems to climate change mitigation is identified in the careful management of nutrients and, hence, the reduction of N2O emissions from soils. The application of biotechnological approaches, developing and implementation of new “omics” technologies and infrastructure construction on a high scientific level will help in building a strategy to overcome the damage of Climate Change. Key words: Climate Change, plant breeding, organic agriculture, plant biotechnology, “OMICS” technologies.

The aim of the current study was to optimize the extraction condition of polyphenol compounds with antioxidant properties from leaves of natural grown Vaccinium myrtillus L and Vaccinium vitis-idaea L. The extractions were carried out in ultrasonic bath at 40 ºC for 20 minutes with different solvents (water, 20 % ethanol, 40 % ethanol, 60 % ethanol, 80 % ethanol and 96 % ethanol) and different hydro module of samples and solvents used (1:50, 1:100 and 1:200). The highest total phenol content in the leaves of V. myrtillus L. and V. vitis-idaea L. was found when 40% ethanol extract was used (90.50 ± 0.05 mg GAE/g DW and 96.68 ± 0.05 mg GAE/g DW, respectively). The highest total flavonoid content of the leaves of both species was obtained with 80 % ethanol as extraction solvent. The highest level of total proanthocyanidins were in the 60 % ethanol extract of V. myrtillus L. and in the 80 % ethanol extract of V. vitis-idaea L (13.12 ± 0.11 mg LE/g DW and 24.22 ± 0.21 mg LE/g DW, respectively). The highest ability to scavenge DPPH radicals possessed the 40 % ethanol extracts from the leaves of both species (693.99 ± 4.05 mM TE/g DW for V. myrtillus L. and 1083.18 ± 8.48 mM TE/g DW for V. vitis-idaea L). Data analysis showed that the maximal amount of polyphenols was extracted at a hydro-module of 1: 100. HPLC analysis revealed that the dominant phenolic acid in the leaves of V. myrtillus L was chlorogenic acid (13.45 mg/g DW), while ferulic acid (49.48 mg/g DW) was present at the highest concentration in the leaves of V. vitis-idaea L.

Arnica montana L. (Asteraceae) is an economically important herb that contains numerous valuable biologically active compounds accumulated in various parts of the plant. The effects of carbon sources (sucrose, maltose, and glucose) at different concentrations (1%, 3%, 5%, 7%, and 9%) on growth were studied and GC-MS based metabolite profiling of A. montana hairy roots was conducted. The optimal growth and biomass accumulation of transformed roots were observed on an MS nutrient medium containing 3% or 5% sucrose. GC-MS analysis of hairy roots of A. montana showed the presence of 48 compounds in polar fractions and 22 compounds in apolar fractions belonging to different classes of metabolites: flavones, phenolic acids, organic acids, fatty acids, amino acids, sugars, sugar alcohols, hydrocarbons etc. Among the various metabolites identified, only the sugars and sugar alcohols were influenced by the concentration of the respective carbon sources in the nutrient medium.

Essential oils and hydrosols of two cultivars of muscadine grapes (Muscadinia rotundifolia (Michx.) Small.) were obtained by hydro-distillation of flowers and berry skins. Twenty-three volatile compounds were identified in essential oils from the muscadine flowers, and twenty volatiles in their corresponding hydrosols. The composition of volatiles in berry skins differed significantly from that of the vine flowers. The antioxidant potential of investigated essential oils and hydrosols was evaluated using five in vitro assays: DPPH (2,2-diphenyl-1-picrylhydrazyl) method, TEAC (Trolox equivalent antioxidant capacity), FRAP (Ferric reducing antioxidant power), CUPRAC (cupric ion reducing antioxidant capacity), and NO (nitric oxide radical scavenging assay). The essential oils from the flowers of both cultivars showed the strongest antioxidant power, whereas the hydrosols were the significantly less active. All investigated essential oils showed very weak antibacterial activities against Bacillus cereus, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. However, the essential oils from the flowers of both cultivars showed moderate antifungal activities against Candida albicans, which were stronger for the oil from “Carlos” (white muscadine cultivar). To the best of our knowledge, this is the first report on obtaining and characterizing essential oils and hydrosols from muscadine grapes. This study demonstrated the variations in aromatic compounds accumulated in flowers and mature berry skins of muscadine grapes, and evaluated their possible antioxidant and antimicrobial activities. The presented results will be the base for future research, focused on a better understanding of the molecular and regulatory mechanisms involved in aromatic compound biosynthesis and accumulation in muscadine grapes.

Leaves of 22 accessions of the Bulgarian raspberry (Rubus L.) germplasm collection – five Bulgarian and seven foreign cultivars, eight elite clones and two wild species, Rubus occidentalis L. and Rubus odoratus L., were analysed for phenolic constituents. The quantitative determination of caffeic (1), p-coumaric (2) and ferulic (3) acids, hyperoside (4), tiliroside (5), and isoquercitrin (6) was performed by RP-HPLC using linear gradient elution and UV detection at 254 and 310 nm. The detection limits ranged from 0.23 mg/ml (4 )t o 0.55 mg/ml (2). Caffeic acid was the dominant phenolic acid in the majority of the samples being present in amounts between 0.05 � 0.01 mg/g dry weight in R. occidentalis and 1.43 � 0.06 mg/g in the cultivars. The highest content of flavonols was found in the Bulgarian raspberry elite clones: 1.70 � 0.002 mg/g (6), 0.60 � 0.004 mg/g (5) and 0.97 � 0.01 mg/g (4). Data were analyzed by hierarchical clustering (HC) and principal component analysis (PCA). The HC and PCA scoring plot showed that the samples could be classified into three clusters. Cluster C1 comprised cultivars characterized by high content of phenolic acids (1–3). Seven cultivars and the wild species R. occidentalis formed the cluster C2 presenting low content of phenolic compounds. Cluster C3 grouped the Bulgarian raspberry elite clones and R. odoratus sharing the highest content of flavonols (5 and 6). The Bulgarian elite clone E23617 displayed the highest content of the studied phenolic derivatives. The PCA loading plot showed that 1 can be used to distinguish between different raspberry varieties.

Summer snowflake/Leucojum aestivum L./is a medical bulbous plant which belongs to family Amarillydaceae. It is wild-spread as natural populations in Bulgaria and a valuable source of galantamine. A few virus and fungal diseases have been described on plants of family Amaryllidaceae but only fungal pathogens on summer snowflake. This paper is the first report of Serratia plymuthica and Stenotrophomonas maltophilia as causal agents of bulb decay of summer snowflake. To our knowledge, this is also the second report of phytopathogenic isolates of the identified two species.The bacteria were isolated from plants in the region of Tutrakan (Bulgaria) close to the Danube River where pathogenic changes in plant tissues of bulbs of summer snowflake were detected in 2006. Pathogenicity to the natural plant host was confirmed by artificial inoculation. Identification was carried out by biochemical properties and sequence analysis. New hosts upon artificial inoculation of S. maltophilia and S. plymuthica are o...

Rhizogenic callus of Salvia scabiosifolia Lam. formed after transformation with Agrobacterium rhizogenes ATCC 15834 was isolated and investigated for triterpenes production. The rhizogenic callus w...

ABTRACTOver the past 10 years, researchers have become increasingly interested in polyphenols. The main reason for this interest is the recognition of the antioxidant properties of polyphenols, their great abundance in our diet, and their probable role in the prevention of various diseases associated with oxidative stress, such as cancer, cardiovascular and neurodegenerative diseases (51). Furthermore, polyphenols, which constitute the active substances found in many plants, modulate the activity of a wide range of enzymes and cell receptors (1). In this way, in addition to having antioxidant properties, polyphenols have several other specific biological actions that are poorly understood.Not all polyphenols are absorbed with equal efficiency. Knowledge of the bioavailability and metabolism of the various polyphenols is necessary in order to evaluate their biological activity within target tissues. The types and distribution of polyphenols in foods and the bioavailability of polyphenols are the topics of ...

The international tendency for growing and production of small fruits shows a permanent increasing. Bulgaria is a traditional important producer of small berries in Europe. A large variety of small fruit products are wide spread and typical for Bulgarian nutriment. Aside with the growing demand in production of small fruit, there is an obvious tendency in food quality, breeding and technology requirements improvement. Breeding purposes comprise improvement of many traits, but selection of disease resistant cultivars, with higher yield and improved consumers properties such as fruit color, shape, smell and transportation ability are among the most important tasks. Recent progress in molecular analyses and agriculture biotechnologies has enormous impact on selection, technology, testing, preservation and processing of agricultural products. Metabolomics assay as a new dimension in these studies and practice focuses the attention on the biochemical contents of cells and tissues, and has a rapidly growing signifi cance in knowledge of small fruits value for human health. Berry fruits are very rich sources of bioactive compounds as phenolics and organic acid. Bioactive berry compounds, their characterization and utilization in functional foods and clinical assessment of antimicrobial properties for human health are among the major targets of contemporary research. Phenolic compounds in berries inhibit the growth of range of human pathogens. Especially raspberry, strawberry, cranberry, crowberries showed evidence of antimicrobial effects against bacterial pathogens as Salmonella and Staphylococcus. The evaluation of small fruit genetic resources for the presence of bioactive compounds and their properties as natural agents is of doubtless signifi cance and will be with great benefi t for breeders, food and pharmaceutical industry.

The genus Rubus contains a large number of highly variably and heterogeneous species, which occur in all parts of the world except the desert regions. Turkey has notable wild grown Rubus populations and these fruits are a traditional part of Turkish diet. The leaves and fruits of wild Rubus forms are used for medicinal purposes in Turkey as well. In the present study we attempted to characterize 11 wild selections and 1 well known red raspberry cultivar, Heritage by using fluorescent dyed AFLP markers and capillary electrophoresis. Four primer combinations generated a total of 259 bands, 182 (69.0%) of which were polymorphic. The higher genetic similarity was found between AH2 and AH3 genotypes. UPGMA clustering of the accessions showed seven major groups. The cultivar Heritage independently formed its own group. AFLP profiles suggesting that the wild genotypes might be derived from seedlings rather than through clonally offshoot propagation.

Bulgarian raspberry germplasm collection contains both Bulgarian and foreign va- rieties with valuable agronomic characters of commercial value. At present, mainly Bul- garian raspberry varieties are grown in Bulgaria with qualities famous in other European countries (Romania, Hungary, Poland, Russia etc.). The study of genetic diversity in the Bulgarian raspberry germplasm collection is mainly based on morphological descrip- tions. Morphological characteristics are important tools but they are not enough for evaluation of the genetic diversity due to the influence of environmental factors and low number of loci studied. The application of molecular markers should give an enable breeders to expand genetic diversity in breeding material and allow selection of new per- spective forms in future breeding programmes. In this article we present our initial study on the assessment of genetic diversity among Bulgarian and foreign raspberry accessions using Simple Sequence Repeat (SSRs) markers. Sixteen raspberry cultivars (six Bulgarian, eight foreign and two wild accessions) were screened for their polymorphism at four loci using polymorphic microsatellite markers. The value determined for genetic variation demonstrated a high genetic diversity in the Bulgarian raspberry germplasm collection. Genetic diversity (GD) calculated from SSR data ranged from 0,740 to 0,887 with a mean GD- 0.839.

GC-MS analysis of alkaloid profiles of five Fumaria species, naturally grown in Bulgaria (F. officinalis, F. thuretii, F. kralikii, F. rostellata and F. schrammii) and analysis of acetylcholinesterase inhibitory activity of alkaloid extracts were performed. Fourteen isoquinoline alkaloids were identified, with the principle ones being protopine, cryptopine, sinactine, parfumine, fumariline, fumarophycine, and fumaritine. Protopine contents, defined by HPLC analysis varied between 210.6 ± 8.8 μg/g DW (F. schrammii) and 334.5 ± 7.1 μg/g DW. (F. rostellata). While all of the investigated alkaloid extracts significantly inhibited acetylcholinesterase activity, the F. kralikii demonstrated the highest level of inhibition (IC50 0.13 ± 0.01 mg extract/mL).

Wild berry species are known to exhibit a wide range of pharmacological activities. They have long been traditionally applied for their antiseptic, antimicrobial, cardioprotective and antioxidant properties. The aim of the present study is to reveal the potential for selective antiviral activity of total methanol extracts, as well as that of the anthocyanins and the non-anthocyanins from the following wild berries picked in Bulgaria: strawberry (Fragaria vesca L.) and raspberry (Rubus idaeus L.) of the Rosaceae plant family, and bilberry (Vaccinium myrtillis L.) and lingonberry (Vaccinium vitis-idaea L) of the Ericaceae. The antiviral effect has been tested against viruses that are important human pathogens and for which chemotherapy and/or chemoprophylaxis is indicated, namely poliovirus type 1 (PV-1) and coxsackievirus B1 (CV-B1) from the Picornaviridae virus family, human respiratory syncytial virus A2 (HRSV-A2) from the Paramyxoviridae and influenza virus A/H3N2 of Orthomyxoviridae. Wild berry fruits are freeze-dried and ground, then total methanol extracts are prepared. Further the extracts are fractioned by solid phase extraction and the non-anthocyanin and anthocyanin fractions are eluted. The in vitro antiviral effect is examined by the virus cytopathic effect (CPE) inhibition test. The results reveal that the total extracts of all tested berry fruits inhibit the replication of CV-B1 and influenza A virus. CV-B1 is inhibited to the highest degree by both bilberry and strawberry, as well as by lingonberry total extracts, and influenza A by bilberry and strawberry extracts. Anthocyanin fractions of all wild berries strongly inhibit the replication of influenza virus A/H3N2. Given the obtained results it is concluded that wild berry species are a valuable resource of antiviral substances and the present study should serve as a basis for further detailed research on the matter.

Hairy root cultures of Salvia tomentosa were initiated by transformation with Agrobacterium rhizogenes. To prevent necrosis in the explants and to protect young hairy roots, Amberlite XAD-4 resin, in combination with a temporary immersion cultivation system, was applied. HPLC analyzes showed that the resin adsorbed more than 93% of the released phenolic acids and 100% of the released flavonoids. The decreased content of the released phenolics significantly reduced their destructive effects on the plant tissues, prevented, and speeded up the appearance of hairy roots.

Wild berry species are known to exhibit a wide range of pharmacological activities. They have long been traditionally applied for their antiseptic, antimicrobial, cardioprotective and antioxidant properties. The aim of the present study is to reveal the potential for selective antiviral activity of total methanol extracts, as well as that of the anthocyanins and the non-anthocyanins from the following wild berries picked in Bulgaria: strawberry ( Fragaria vesca L.) and raspberry ( Rubus idaeus L.) of the Rosaceae plant family, and bilberry ( Vaccinium myrtillis L.) and lingonberry ( Vaccinium vitis-idaea L) of the Ericaceae. The antiviral effect has been tested against viruses that are important human pathogens and for which chemotherapy and/or chemoprophylaxis is indicated, namely poliovirus type 1 (PV-1) and coxsackievirus B1 (CV-B1) from the Picornaviridae virus family, human respiratory syncytial virus A2 (HRSV-A2) from the Paramyxoviridae and influenza virus A/H3N2 of Orthomyxoviridae. Wild berry fruits are freeze-dried and ground, then total methanol extracts are prepared. Further the extracts are fractioned by solid phase extraction and the non-anthocyanin and anthocyanin fractions are eluted. The in vitro antiviral effect is examined by the virus cytopathic effect (CPE) inhibition test. The results reveal that the total extracts of all tested berry fruits inhibit the replication of CV-B1 and influenza A virus. CV-B1 is inhibited to the highest degree by both bilberry and strawberry, as well as by lingonberry total extracts, and influenza A by bilberry and strawberry extracts. Anthocyanin fractions of all wild berries strongly inhibit the replication of influenza virus A/H3N2. Given the obtained results it is concluded that wild berry species are a valuable resource of antiviral substances and the present study should serve as a basis for further detailed research on the matter.

This review focused on application a new biotechnology and molecular methods for preservation, evaluation and utilization of Plant Genetic Resources. The assessment of Bulgarian Small Fruits collections take up very important place in agro-horticultural crops with great significance for breeders. A number of strategies for collection, preservation and sustained use of genetic resources for agricultural productivity and food security were applicable. In vitro techniques offer the opportunity for in vitro collecting, rapid propagation, medium and long-term storage of germplasm and its distribution. The application of molecular markers will allow to selection of new perspective forms and including in future breeding programme.